Preterm labour prevention: composition and characterisation of the predominate KV7 channel subunits in human uterine tissue and smooth muscle cells

尽管自然早产是世界范围内围产儿发病率和死亡率的主要因素，但它不能被准确地预测、预防或治疗。我们对子宫收缩控制的研究使我们提出了由KCNQ2-5基因编码的钾通道(KV7通道)作为潜在的药物靶点来抑制早产子宫收缩和潜在的免疫细胞激活。只有特定的KCNQ1-5和附属的KCNE1-5在妊娠妇女的子宫组织(子宫肌层)中有功能表达。体外药理激活KV7通道抑制妊娠人和小鼠子宫肌层收缩，其中ML213(KV7.4/KV7.4、KV7.2和Kv7.5优先激活剂)最有效。KV7通道激活剂在一个实验模型中延迟早产。这为人类子宫肌层中具有功能的KV7通道提供了必要的“原则性证据”数据，并突出了KV7通道激活剂抑制子宫收缩和早产的潜力。该PHD项目的目的是确定人类子宫KV7通道的特定结构及其与KCNE蛋白的相互作用，以及KV7.4通道在人类子宫肌层组织和细胞中的调节和运输。第二个目的是确定KV7表达的子宫细胞类型差异，以了解潜在的药理学影响(例如免疫细胞)。将使用各种成像和分子技术，包括使用超分辨率共聚焦成像、多光子显微镜、结构照明成像、基因敲除和过度表达以及RNAseq。该项目可以灵活地根据候选人的兴趣/技能集，更多地关注蛋白质结构、细胞调节和基因操纵或膜生物物理方法。将在两位主管设备齐全的研究设施/实验室提供培训。学生将有机会接触KCL CORE和BRC的设施(例如尼康成像中心和BRC流式细胞术和基因组学平台)，相关的个人发展培训，以及杂志俱乐部和研讨会。

Despite spontaneous preterm birth being a major contributor to perinatal morbidity and mortality worldwide, it cannot be accurately predicted or prevented or treated. Our research into the control of uterine contractility has led us to propose potassium channels encoded by KCNQ2-5 genes (KV7 channels) as a potential drug target to inhibit preterm uterine contractions and potentially immune cell activation. Only specific KCNQ1-5 and accessory KCNE1-5 are functionally expressed in pregnant human uterine tissue (myometrium) from women at term and in labour. Pharmacological activation of Kv7 channels inhibit pregnant human and mouse myometrium contractions in vitro, with ML213 (a KV7.4/ KV7.4, KV7.2and Kv7.5 preferential activator) being the most effective. Kv7 channel activators delay preterm birth in an experimental model. This provides essential 'proof of principle' data for functional KV7 channels in human myometrium and highlights the potential of KV7 channel activators to inhibit uterine contractions and preterm delivery. The aim of this PhD project is to determine the specific architecture of the human uterine KV7 channel and its interactions with KCNE proteins as well as aspects of KV7.4 channel regulation and trafficking in human myometrium tissue and cells. A secondary aim is to determine uterine cell type differences in Kv7 expression in order to understand potential pharmacological impact (e.g. immune cells). A variety of imaging and molecular techniques will be used including using super resolution confocal imaging, multiphoton microscopy, structural illumination imaging, gene knockdown and overexpression, and RNAseq. The project has flexibility to focus more on protein structure, cellular regulation and gen e manipulation or membrane biophysics approaches depending on candidate interest/skill set. Training will be provided in both supervisors' well-equipped research facilities/laboratories. The student will have access to KCL core and BRC facilities (e.g. Nikon Imaging Centre and BRC Flow cytometry and genomics platforms), relevant personal development training, and journal clubs and seminars.